CAJ | 학술논문

目的分析不同浓度的灵芪蠲肝胶囊含药血清对血小板源性生长因子(PDGF)诱导的大鼠肝星状细胞-T6 (HSC-T6)增殖和酪氨酸激酶2(JAK2)、磷酸化的信号传导与转录活化因子3(p-STAT3)蛋白量表达的影响及可能的调控机制. 方法 SD大鼠25只,随机分为5组:正常血清对照组(A组)、复方鳖甲软肝片(B组)、灵芪蠲肝胶囊小剂量组(C1组)、灵芪蠲肝胶囊中剂量组(C2组)、灵芪蠲肝胶囊大剂量组(C3组),药物灌胃7d,经腹主动脉采血,制备含药血清.将各组含药血清分别作用于10 ng/ml PDGF刺激的HSC-T6,24、48、72 h后用CCK-8法测定各组HSC-T6的吸光度值,免疫印迹法检测24 h时各组细胞中JAK2、p-STAT3蛋白量的表达.各组重复测量的数据采用重复测量的方差分析,各组间比较用单因素方差分析,多样本均数两两比较用SNK-q检验.结果与A组比较,C2、C3组SD大鼠药物血清在24、48、72h均可明显抑制HSC-T6的增殖,根据重复测量的方差分析,F=36.292,P值均＜0.05,差异均有统计学意义.C1组24 h吸光度值为1.522±0.128,与A组24h的吸光度值1.550±0.065比较,P＞0.05,差异无统计学意义;C1组48、72 h可抑制HSC-T6增殖,吸光度值分别为1.153±0.183、0.753±0.210,与A组48、72 h的吸光度值1.311±0.650和1.039±0.103比较,F=36.292,p值均＜0.05,差异均有统计学意义.与A组比较,24 h时C1、C2、C3各组HSC-T6中JAK2、p-STAT3蛋白表达量均明显减少.结论灵芪蠲肝胶囊能抑制PDGF诱导的HSC-T6增殖,且作用呈剂量依赖性；同时能降低JAK2、p-STAT3在活化的HSC-T6中的表达,阻碍JAK/STAT信号通路的传导,发挥抗肝纤维化作用.
목적 분석불동농도적령기견간효낭함약혈청대혈소판원성생장인자(PDGF)유도적대서간성상세포-T6 (HSC-T6)증식화락안산격매2(JAK2)、린산화적신호전도여전록활화인자3(p-STAT3)단백량표체적영향급가능적조공궤제. 방법 SD대서25지,수궤분위5조:정상혈청대조조(A조)、복방별갑연간편(B조)、령기견간효낭소제량조(C1조)、령기견간효낭중제량조(C2조)、령기견간효낭대제량조(C3조),약물관위7d,경복주동맥채혈,제비함약혈청.장각조함약혈청분별작용우10 ng/ml PDGF자격적HSC-T6,24、48、72 h후용CCK-8법측정각조HSC-T6적흡광도치,면역인적법검측24 h시각조세포중JAK2、p-STAT3단백량적표체.각조중복측량적수거채용중복측량적방차분석,각조간비교용단인소방차분석,다양본균수량량비교용SNK-q검험.결과 여A조비교,C2、C3조SD대서약물혈청재24、48、72h균가명현억제HSC-T6적증식,근거중복측량적방차분석,F=36.292,P치균＜0.05,차이균유통계학의의.C1조24 h흡광도치위1.522±0.128,여A조24h적흡광도치1.550±0.065비교,P＞0.05,차이무통계학의의;C1조48、72 h가억제HSC-T6증식,흡광도치분별위1.153±0.183、0.753±0.210,여A조48、72 h적흡광도치1.311±0.650화1.039±0.103비교,F=36.292,p치균＜0.05,차이균유통계학의의.여A조비교,24 h시C1、C2、C3각조HSC-T6중JAK2、p-STAT3단백표체량균명현감소.결론 령기견간효낭능억제PDGF유도적HSC-T6증식,차작용정제량의뢰성；동시능강저JAK2、p-STAT3재활화적HSC-T6중적표체,조애JAK/STAT신호통로적전도,발휘항간섬유화작용.
Objective To analyze the effects of Ling Qi Juan Gan capsule drug-containing serum at different concentrations on the platelet-derived growth factor (PDGF)-induced proliferative capabilities of and JAK2 and p-STAT3 protein expression in hepatic stellate cells (HSC) using an in vitro system.Methods Twenty-five Sprague-Dawley rats were randomly divided into five equal groups for intragastric administration of physiological saline (10 ml/kg; group A),Fufang Biejia Ruangan tablet solution (1.5 g/kg; group B),or Ling Qi Juan Gan capsule solution at low dose (2.125 g/kg; group C1),mid dose (4.25 g/kg; group C2),or high dose (8.5 g/kg; group C3).The post-administration serum isolated from each rat (200 ml/L) was used to treat the HSC-T6 cell line following induction by PDGF (10 ng/ml).At 24,48 and 72 h post-exposure,the cells' proliferation was measured using the Cell Counting Kit-8 (CCK-8) colorimetric assay.In addition,at 24 h post-exposure the expression of JAK2 and p-STAT3 was measured by western blotting (expressed as grey scale intensity).Multiple group comparison of repeated measures data was made by one-way ANOVA with Student-Newman-Keuls post hoc test.Results Compared to group A,groups C2 and C3 had significantly higher inhibited proliferation at all post-exposure time points examined (24 h:A =1.550 ± 0.065,C2 =1.335 ± 0.106,C3 =1.241 ± 0.205; 48 h:A =1.311 ± 0.650,C2 =1.090 ± 0.106,C3 =0.909 ± 0.191; 72 h:A =1.039 ± 0.103,C2 =0.719 ± 0.116,C3 =0.641 ± 0.110,F =36.292,all P ＜ 0.05); in contrast,compared to group A,group C1 showed no inhibition of proliferation at 24 h (1.522 ± 0.128,P =0.717) but showed significantly higher inhibition of proliferation at 48 h and 72 h (1.153 ± 0.183 and 0.753 ± 0.210,respectively,F =36.292,P ＜0.05).Compared with group A,all Ling Qi Juan Gan capsule-containing serum-treated groups showed significantly lower expression of both JAK2 (A =1.605 ± 0.024 vs.C1 =1.170 ± 0.042,C2 =0.842 ± 0.036,C3 =0.555 ± 0.036,F=43.091) and p-STAT3 (A =1.401 ± 0.030 vs.C1 =1.229 ± 0.025,C2 =0.668 ±0.034,C3 =0.630 ± 0.026,F =78.426,all P ＜ 0.01).Conclusion Ling Qi Juan Gan capsule drug-containing serum can inhibit the proliferation of HSC-T6 cells in a dose-dependent manner and cause an overall decrease in the expression of JAK2 and p-STAT3 in activated HSC,thereby leading to a suppression of the JAK/STAT signal transduction pathway.